Discocleidion sufescens Pax et Hoffm. extract, preparation method thereof and purpose thereof
A technology for extracting and wrapping leaves, which is applied in the field of heaven and can solve problems such as separation of effective parts of Alzheimer's disease that has not been seen
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Embodiment 1
[0016] Pseudomonas herb (10.0Kg), dried, crushed and extracted with 10 times, 8 times and 8 times of 95% pharmaceutical grade ethanol respectively for 3 times, each time for 2 hours, the extract was filtered with gauze and combined Three extracts; low temperature (<80°C) to recover ethanol under reduced pressure until there is no alcohol smell to obtain ethanol extract; suspend the ethanol extract with distilled water, and use petroleum ether 5000mL×5 times, ethyl acetate 4000mL×5 times, water Saturated n-butanol 5000mL×3 extractions; the aqueous solution after the above-mentioned petroleum ether, ethyl acetate and water-saturated n-butanol extracts was concentrated under reduced pressure and low temperature (<80°C) to recover the organic solvent until there was no solvent smell, and obtained Active part extract 1.5Kg. 95% ethanol extract, petroleum ether extract, ethyl acetate extract, n-butanol extract, and active site extract were respectively used for nerve cell activity s...
Embodiment 2
[0018] Example 2: Structural identification of the active site
[0019] 100g of the active part of the pseudo-capsule, mix the sample with silica gel (0.063-0.200mm, 250g), put it on a silica gel column (0.060-0.200mm) for chromatographic separation, and dichloromethane-methanol-water (100:0:0-100 : 30:10) gradient elution, each 200mL as a fraction, a total of 100 fractions; dichloromethane-methanol-water (100:10:2) fraction through silica gel column (0.040-0.063mm), ODS and Sephadex LH- 20 column chromatographic separation to obtain compounds Ⅰ (25mg), Ⅱ (30mg); dichloromethane-methanol-water (100:10:100) was separated by silica gel column (0.040-0.063mm) and ODS column chromatography, and separated Compound Ⅲ (30 mg) was obtained; dichloromethane-methanol-water (100:20:5) part was separated by silica gel (0.040-0.063mm) and ODS repeated column chromatography to obtain compound Ⅳ (25 mg), Ⅴ (30 mg) , VI (25mg).
[0020] The structures of compound Ⅰ~compound Ⅵ were identif...
Embodiment 3
[0025] Example 3: Preliminary research data on the anti-senile dementia effect of the active site
[0026] Take 95% ethanol extract, petroleum ether extract, ethyl acetate extract, n-butanol extract, and active site extract respectively for nerve cell activity screening. The results show that 95% ethanol extract and active site extract are all Has a strong NGF-inducing activity.
[0027] 1. Principle and method
[0028] Using the nerve cell in vitro model test, using PC-12 cells, to detect the NGF-inducing activity of each part.
[0029] (1) PC 12 cells are cell lines differentiated from rat adrenal medulla pheochromocytoma, which have general characteristics of neuroendocrine cells, and are now widely used in neurophysiological and neuropharmacological research.
[0030] When cultured with very low concentration of NGF, PC 12 cells can be induced to differentiate into NGF-dependent dopaminergic neurons.
[0031] (2) LDH (Lactic dehydrogenase, lactate dehydrogenase) is us...
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